Wadjemup comparison
2023-10-09
This is the same notebook as 100_species, but we look at the wadjemup/rottnest queryset. This should be slightly harder. # Setup
knitr::opts_chunk$set(warning = FALSE)
source(here::here('code', 'helpers.R'))
library(tidyverse)
library(forcats)
library(cowplot)
library(ggupset)
library(RColorBrewer)
library(patchwork)
library(vegan)
library(pheatmap)
library(broom)data <- targets::tar_read(merged_all_results)
# rename BLAST to BLAST97 to differentiate from BLAST100 (percentage identity in both cases)
data <- data |> mutate(Type = str_replace(Type, '^BLAST$', 'BLAST97'))
truth <- targets::tar_read(truth_set_data)table(data$Type)##
## BLAST100 BLAST97 CustomNBC DADA2Spec DADA2Tax Kraken_0.0
## 13912 21093 155700 157104 157104 104880
## Kraken_0.05 Kraken_0.1 Metabuli MMSeqs2_100 MMSeqs2_97 Mothur
## 104880 104880 73217 109368 109368 152892
## Qiime2 VSEARCH
## 23275 157104Let’s remove the >0.2 Kraken runs, those are too strict
data <- data |> filter(!Type %in% c('Kraken_0.2', 'Kraken_0.3', 'Kraken_0.4', 'Kraken_0.5', 'Kraken_0.6', 'Kraken_0.7', 'Kraken_0.8', 'Kraken_0.9'))Made a mistake- Metabuli’s and TNT’s databases is misspelled
data <- data |> mutate(Subject = str_replace_all(Subject, pattern = '_ref.fasta', replacement=''))
data <- data |> mutate(Subject = str_replace_all(Subject, pattern = 'final.csv', replacement = 'final.fasta'))table(data$Query)##
## KWest_16S_PooledTrue.fa
## 366180
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa
## 100033
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa
## 99769
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa
## 97434
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa
## 97254
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 101106
## make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa
## 88422
## make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa
## 89094
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa
## 24355
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa
## 26746
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 26505
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa
## 102811
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa
## 109807
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 115261Wadjemup species
Check 12S results
table(data$Subject)##
## 12s_v010_final.fasta
## 16590
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 16175
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 15914
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 15918
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 16012
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 16089
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 16203
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 15855
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 15966
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 16261
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 16047
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 16366
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 16276
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 16170
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 16336
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 16075
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 16297
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 15985
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 16148
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 16105
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 16282
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 15716
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 15455
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 15781
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 15799
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 15581
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 15704
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 15553
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 15589
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 15583
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 15626
## 12s_v010_finalforTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## 12S_v10_HmmCut.fasta
## 8714
## 16S_v04_final.fasta
## 17674
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 16374
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 16360
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 16735
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 16567
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 16776
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 16689
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 16330
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 16841
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 16165
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 16285
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 17050
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 16867
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 17165
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 16830
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 17045
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 17369
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 16953
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 17104
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 17262
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 17159
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 16172
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 16327
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 16294
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 16191
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 16435
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 16174
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 15983
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 16145
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 16010
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 16118
## 16S_v04_finalforTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_HmmCut.fasta
## 9590
## c01_v03_final.fasta
## 12609
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 12286
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 12907
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 12333
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 12919
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 12107
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 12272
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 12223
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 12673
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 12229
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 12925
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 12455
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 12383
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 12356
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 12334
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 12420
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 12446
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 12329
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 12419
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 12572
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 12499
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 12989
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 12948
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 12809
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 12053
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 12010
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 12513
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 12803
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 12641
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 11991
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 12002
## c01_v03_finalforTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_HmmCut.fasta
## 6792table(data$Subject)##
## 12s_v010_final.fasta
## 16590
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 16175
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 15914
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 15918
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 16012
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 16089
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 16203
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 15855
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 15966
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 16261
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 16047
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 16366
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 16276
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 16170
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 16336
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 16075
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 16297
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 15985
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 16148
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 16105
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 16282
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 15716
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 15455
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 15781
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 15799
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 15581
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 15704
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 15553
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 15589
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 15583
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 15626
## 12s_v010_finalforTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## 12S_v10_HmmCut.fasta
## 8714
## 16S_v04_final.fasta
## 17674
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 16374
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 16360
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 16735
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 16567
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 16776
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 16689
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 16330
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 16841
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 16165
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 16285
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 17050
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 16867
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 17165
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 16830
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 17045
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 17369
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 16953
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 17104
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 17262
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 17159
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 16172
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 16327
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 16294
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 16191
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 16435
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 16174
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 15983
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 16145
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 16010
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 16118
## 16S_v04_finalforTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## 16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## 16S_v04_HmmCut.fasta
## 9590
## c01_v03_final.fasta
## 12609
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 12286
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 12907
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 12333
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 12919
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 12107
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 12272
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 12223
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 12673
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 12229
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 12925
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 12455
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 12383
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 12356
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 12334
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 12420
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 12446
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 12329
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 12419
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 12572
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 12499
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 12989
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 12948
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 12809
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 12053
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 12010
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 12513
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 12803
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 12641
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 11991
## c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 12002
## c01_v03_finalforTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta
## 240
## c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta
## 240
## c01_v03_HmmCut.fasta
## 6792twelveS_data <- data |> filter(Subject == '12s_v010_final.fasta')
sixteenS_data <- data |> filter(Subject == '16S_v04_final.fasta')
table(twelveS_data$Query)##
## KWest_16S_PooledTrue.fa
## 4263
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa
## 1321
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa
## 1321
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa
## 1095
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa
## 1095
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 968
## make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa
## 990
## make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa
## 1000
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa
## 316
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa
## 299
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 255
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa
## 1343
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa
## 1226
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 1098table(sixteenS_data$Query)##
## KWest_16S_PooledTrue.fa
## 4928
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa
## 1230
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa
## 1194
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa
## 1306
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa
## 1306
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 968
## make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa
## 990
## make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa
## 1000
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa
## 307
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa
## 341
## make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 255
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa
## 1274
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa
## 1477
## make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa
## 1098table(sixteenS_data$Subject)##
## 16S_v04_final.fasta
## 17674twelveS_data_vs_12S_100 <- twelveS_data |>
filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa')
sixteenS_data_vs_16S_100 <- sixteenS_data |>
filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa' )twelveS_data_vs_12S_100 |> select(Type, species) |> filter(species != 'dropped' &
!is.na(species)) |>
group_by(Type) |> count(species) |> summarise(n = n()) |>
ggplot(aes(x = Type, y = n, fill = n)) + geom_col() + coord_flip() +
theme_minimal() +
ylab('Count') +
ggtitle('12S: Species-level hits per classifier')
twelveS_data_vs_12S_100 |> select(Type, genus) |> filter(genus != 'dropped' &
!is.na(genus)) |>
group_by(Type) |> count(genus) |> summarise(n = n()) |>
ggplot(aes(x = Type, y = n, fill = n)) + geom_col() + coord_flip() +
theme_minimal() +
ylab('Count') +
ggtitle('12S: Genus-level hits per classifier')
twelveS_truth <- truth |> filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa') |> select(OTU, family, species) |> rename(True_OTU = OTU, True_family = family, True_species = species)
head(twelveS_truth)## # A tibble: 6 × 3
## True_OTU True_family True_species
## <chr> <chr> <chr>
## 1 ASV_1 Syngnathidae Phyllopteryx taeniolatus
## 2 ASV_2 Syngnathidae Phycodurus eques
## 3 ASV_3 Syngnathidae Phyllopteryx taeniolatus
## 4 ASV_4 Labridae dropped
## 5 ASV_5 Alopiidae Isurus oxyrinchus
## 6 ASV_6 Carangidae Seriola lalanditwelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
mutate(Correct = True_species == species) |>
filter(species != 'dropped' & !is.na(species)) |>
group_by(Type) |> count(Correct) |>
ggplot(aes(x = fct_rev(fct_reorder2(Type, Correct, n)), fill = Correct, y = n))+ geom_col() +
coord_flip() + theme_minimal() + xlab('Type') +
ggtitle('12S: Correct and incorrect species-level classifications (absolute)') +
scale_fill_manual(values = c("#E69F00", "#56B4E9", "#009E73",
"#F0E442", "#0072B2", "#D55E00", "#CC79A7"))
cols <- c('Correct species' = "#009E73", 'Correct genus'="#56B4E9", 'Correct family' = "#0072B2", 'Incorrect family' = "#E69F00", 'Incorrect genus'="#F0E442", 'Incorrect species'="#D55E00", 'No hit'= "#CC79A7")twelve_s_relative_plot <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type) |>
count(CorrectSpecies) |>
mutate(total = sum(n, na.rm=TRUE)) |>
mutate(missing = 102 - total) |>
group_modify(~ add_row(.x)) |>
group_modify(~ {
.x |> mutate(new_col= max(missing, na.rm=TRUE)) |>
mutate(n = case_when(is.na(CorrectSpecies) & is.na(missing) ~ new_col,
TRUE ~ n)) |>
select(-new_col)
} ) |>
mutate(total = 102) |>
mutate(perc = n / total * 100) |>
mutate(CorrectSpecies = replace_na(CorrectSpecies, 'No hit')) |>
mutate(CorrectSpecies = factor(CorrectSpecies, rev(c('Correct species', 'Correct genus', 'Correct family', 'Incorrect family', 'Incorrect genus', 'Incorrect species', 'No hit')))) |>
ggplot(aes(x = fct_rev(fct_reorder2(Type, CorrectSpecies, n)), fill = CorrectSpecies, y = perc))+
geom_col() +
coord_flip() +
theme_minimal() +
ylab('Percentage') + xlab('Type') +
ggtitle('12S: Correct and incorrect species-level classifications (relative)') +
scale_fill_manual(name = 'Outcome', values = cols, breaks=names(cols))
twelve_s_relative_plot
### Calculate Upset-based species sightings
type_list <- twelveS_data_vs_12S_100 |> select(Type, species) |> unique() |> filter(!is.na(species) & species != 'dropped') |>
group_by(species) |>
summarize('Type' = list(Type))
a <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('12S: Shared species') +
ylab('Species')
a
type_list <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
filter(species == True_species) |>
filter(species != 'dropped' & !is.na(species)) |>
select(Type, species) |> unique() |>
group_by(species) |>
summarize('Type' = list(Type))
b <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('12S: Shared correct species') +
ylab('Species')
b
type_list <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
filter(species != True_species) |>
filter(species != 'dropped' & !is.na(species)) |>
select(Type, species) |> unique() |>
group_by(species) |>
summarize('Type' = list(Type))
c <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('12S: Shared incorrect species') +
ylab('Species')
c
a + b + c & ylim(c(0, 30)) &
theme(
# Hide panel borders and remove grid lines
panel.border = element_blank(),
panel.grid.major = element_blank(),
panel.background = element_blank(),
panel.grid.minor = element_blank(),
#panel.grid.major.y = element_line(),
# Change axis line
axis.line = element_line(colour = "black")
)
Calculate FP/TP/TN/FN
add_scores <- function(twelveS_data_vs_12S_100_with_MaxTruth, twelveS_truth ) {
twelveS_data_vs_12S_100_with_MaxTruth|> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type) |>
summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
FN = sum(is.na(species) & !is.na(True_species))) |>
mutate(sums = TP + FP + TN + FN) |>
mutate(missing = 102 - sums) |>
mutate(FN = FN + missing) |>
mutate(sums = TP + FP + TN + FN) |>
select(-c(missing, sums))
}scores <- add_scores(twelveS_data_vs_12S_100, twelveS_truth)scores <- scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))
scores## # A tibble: 14 × 10
## # Rowwise:
## Type TP FP TN FN recall precision f1 f0.5 accuracy
## <chr> <int> <int> <int> <dbl> <dbl> <dbl> <dbl> <dbl> <dbl>
## 1 BLAST100 57 2 0 43 0.57 0.966 0.717 0.848 0.559
## 2 BLAST97 59 8 0 35 0.628 0.881 0.733 0.815 0.578
## 3 CustomNBC 53 18 0 31 0.631 0.746 0.684 0.720 0.520
## 4 DADA2Spec 57 3 0 42 0.576 0.95 0.717 0.841 0.559
## 5 DADA2Tax 7 26 0 69 0.0921 0.212 0.128 0.168 0.0686
## 6 Kraken_0.0 60 12 0 30 0.667 0.833 0.741 0.794 0.588
## 7 Kraken_0.05 57 9 0 36 0.613 0.864 0.717 0.798 0.559
## 8 Kraken_0.1 53 5 0 44 0.546 0.914 0.684 0.805 0.520
## 9 MMSeqs2_100 58 2 0 42 0.58 0.967 0.725 0.853 0.569
## 10 MMSeqs2_97 64 7 0 31 0.674 0.901 0.771 0.844 0.627
## 11 Metabuli 65 11 0 26 0.714 0.855 0.778 0.823 0.637
## 12 Mothur 42 22 0 38 0.525 0.656 0.583 0.625 0.412
## 13 Qiime2 31 18 0 53 0.369 0.633 0.466 0.554 0.304
## 14 VSEARCH 52 17 0 33 0.612 0.754 0.675 0.720 0.510twelveS_scoreS_plot <- scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |> ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1)) + theme_minimal_hgrid()+ theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') + ggtitle('12S scores')
twelveS_scoreS_plot
Scores heatmap
Let’s also make a heatmap from that
b <- scores$Type
m <- scores |> select(-Type) |> select(accuracy, recall, precision, f1, f0.5) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
color = colorRampPalette(brewer.pal(n = 7, name =
"RdYlGn"))(100))
b <- scores$Type
m <- scores |> select(-Type) |> select(TP, FP, FN) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
color = colorRampPalette(brewer.pal(n = 7, name =
"RdYlGn"))(100))
Metaclassifier
table(twelveS_data_vs_12S_100$Type)##
## BLAST100 BLAST97 CustomNBC DADA2Spec DADA2Tax Kraken_0.0
## 79 93 102 102 102 102
## Kraken_0.05 Kraken_0.1 Metabuli MMSeqs2_100 MMSeqs2_97 Mothur
## 102 102 102 102 102 102
## Qiime2 VSEARCH
## 49 102First, we count the per-OTU species hits
twelveS_data_vs_12S_100_maxCount <- twelveS_data_vs_12S_100 |>
mutate(species = na_if(species, 'dropped')) |>
filter(!is.na(species)) |>
#filter(! Type %in% c('Mothur', 'VSEARCH', 'Kraken_0.2', 'Qiime2', 'Metabuli', 'NBC', 'BLAST97', 'Kraken_0.0', 'Kraken_0.1')) |>
group_by(Query, Subject, OTU) |>
count(species) |>
# double check the truth
#left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
#mutate(Truth = True_species == species) |>
# pull out the per-group highest n
filter( n > 4) |>
slice_max(n, n=1, with_ties = FALSE) |>
mutate(Type = 'MaxCount', .before = 'Query') |>
select(-n)
twelveS_data_vs_12S_100_maxCount## # A tibble: 75 × 5
## # Groups: Query, Subject, OTU [75]
## Type Query Subject OTU species
## <chr> <chr> <chr> <chr> <chr>
## 1 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_1 Phyllo…
## 2 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Carcha…
## 3 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Isurus…
## 4 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Acanth…
## 5 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Pseudo…
## 6 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Carcha…
## 7 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Parupe…
## 8 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Sphyrn…
## 9 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Parupe…
## 10 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Chaeto…
## # ℹ 65 more rowstwelveS_data_vs_12S_100_with_MaxTruth <- twelveS_data_vs_12S_100 |>
bind_rows(twelveS_data_vs_12S_100_maxCount) #|>
#filter(! Type %in% c('Mothur', 'VSEARCH', 'Kraken_0.2', 'Qiime2', 'Metabuli', 'NBC', 'BLAST97', 'Kraken_0.0', 'Kraken_0.1')) maxTruth_scores <- add_scores(twelveS_data_vs_12S_100_with_MaxTruth, twelveS_truth )maxTruth_scores <- maxTruth_scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))maxTruth_scoreS_plot <- maxTruth_scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |> ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1)) + theme_minimal_hgrid()+ theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') + geom_point() + ggtitle('12S scores')
maxTruth_scoreS_plot
With the Wadjemup data MaxCount performs slightly better! But still not
as good as MMSeqs2….
16S
sixteenS_truth <- truth |> filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa') |> select(OTU, family, species) |> rename(True_OTU = OTU, True_family = family, True_species = species)
tail(sixteenS_truth)## # A tibble: 6 × 3
## True_OTU True_family True_species
## <chr> <chr> <chr>
## 1 ASV_106 Alopiidae Isurus oxyrinchus
## 2 ASV_107 Carangidae Seriola dumerili
## 3 ASV_108 Carcharhinidae Sphyrna lewini
## 4 ASV_109 Scombridae Auxis thazard
## 5 ASV_110 Syngnathidae Phycodurus eques
## 6 ASV_111 Syngnathidae Phyllopteryx taeniolatussixteenS_relative_plot <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type) |>
count(CorrectSpecies) |>
mutate(total = sum(n)) |>
mutate(missing = 112 - total) |>
group_modify(~ add_row(.x)) |>
group_modify(~ {
.x |> mutate(new_col= max(missing, na.rm=TRUE)) |>
mutate(n = case_when(is.na(CorrectSpecies) & is.na(missing) ~ new_col,
TRUE ~ n)) |>
select(-new_col)
} ) |>
mutate(total = 112) |>
mutate(perc = n / total * 100) |>
mutate(CorrectSpecies = replace_na(CorrectSpecies, 'No hit')) |>
mutate(CorrectSpecies = factor(CorrectSpecies, rev(c('Correct species', 'Correct genus', 'Correct family', 'Incorrect family', 'Incorrect genus', 'Incorrect species', 'No hit')))) |>
tidyr::complete(CorrectSpecies, fill = list(n=0, total = 112, missing = NA, perc = 0)) |>
ggplot(aes(x = fct_rev(fct_reorder2(Type, CorrectSpecies, n)), fill = CorrectSpecies, y = perc))+
geom_col() +
coord_flip() +
theme_minimal() +
ylab('Percentage') + xlab('Type') +
ggtitle('16S: Correct and incorrect species-level classifications (relative)') +
scale_fill_manual(name = 'Outcome', values = cols, breaks=names(cols))
sixteenS_relative_plot 
Calculate scores
scores <- add_scores(sixteenS_data_vs_16S_100, sixteenS_truth)
scores## # A tibble: 14 × 5
## Type TP FP TN FN
## <chr> <int> <int> <int> <dbl>
## 1 BLAST100 53 3 0 46
## 2 BLAST97 62 6 2 32
## 3 CustomNBC 56 18 2 26
## 4 DADA2Spec 52 4 2 44
## 5 DADA2Tax 8 40 2 52
## 6 Kraken_0.0 70 16 2 14
## 7 Kraken_0.05 69 11 2 20
## 8 Kraken_0.1 66 8 2 26
## 9 MMSeqs2_100 54 3 2 43
## 10 MMSeqs2_97 69 8 2 23
## 11 Metabuli 69 18 2 13
## 12 Mothur 61 27 2 12
## 13 Qiime2 56 20 1 25
## 14 VSEARCH 68 11 2 21scores <- scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))
scores## # A tibble: 14 × 10
## # Rowwise:
## Type TP FP TN FN recall precision f1 f0.5 accuracy
## <chr> <int> <int> <int> <dbl> <dbl> <dbl> <dbl> <dbl> <dbl>
## 1 BLAST100 53 3 0 46 0.535 0.946 0.684 0.820 0.520
## 2 BLAST97 62 6 2 32 0.660 0.912 0.765 0.847 0.627
## 3 CustomNBC 56 18 2 26 0.683 0.757 0.718 0.741 0.569
## 4 DADA2Spec 52 4 2 44 0.542 0.929 0.684 0.813 0.529
## 5 DADA2Tax 8 40 2 52 0.133 0.167 0.148 0.159 0.0980
## 6 Kraken_0.0 70 16 2 14 0.833 0.814 0.824 0.818 0.706
## 7 Kraken_0.05 69 11 2 20 0.775 0.862 0.817 0.844 0.696
## 8 Kraken_0.1 66 8 2 26 0.717 0.892 0.795 0.851 0.667
## 9 MMSeqs2_100 54 3 2 43 0.557 0.947 0.701 0.831 0.549
## 10 MMSeqs2_97 69 8 2 23 0.75 0.896 0.817 0.862 0.696
## 11 Metabuli 69 18 2 13 0.841 0.793 0.817 0.802 0.696
## 12 Mothur 61 27 2 12 0.836 0.693 0.758 0.718 0.618
## 13 Qiime2 56 20 1 25 0.691 0.737 0.713 0.727 0.559
## 14 VSEARCH 68 11 2 21 0.764 0.861 0.810 0.840 0.686sixteenS_score_plot <- scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |> ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1)) + theme_minimal_hgrid() + theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') + ggtitle('16S scores')
sixteenS_score_plot 
b <- scores$Type
m <- scores |> select(-Type) |> select(accuracy, recall, precision, f1, f0.5) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
color = colorRampPalette(brewer.pal(n = 7, name =
"RdYlGn"))(100))
b <- scores$Type
m <- scores |> select(-Type) |> select(TP, FP, FN) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
color = colorRampPalette(brewer.pal(n = 7, name =
"RdYlGn"))(100))
### Calculate Upset-based species sightings
type_list <- sixteenS_data_vs_16S_100 |> select(Type, species) |> unique() |> filter(!is.na(species) & species != 'dropped') |>
group_by(species) |>
summarize('Type' = list(Type))
a <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('16S: Shared species') +
ylab('Species')
a
type_list <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
filter(species == True_species) |>
filter(species != 'dropped' & !is.na(species)) |>
select(Type, species) |> unique() |>
group_by(species) |>
summarize('Type' = list(Type))
b <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('16S: Shared correct species') +
ylab('Species')
b
type_list <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
filter(species != True_species) |>
filter(species != 'dropped' & !is.na(species)) |>
select(Type, species) |> unique() |>
group_by(species) |>
summarize('Type' = list(Type))
c <- type_list |>
ggplot(aes(x = Type)) +
geom_bar() +
scale_x_upset(n_intersections = 12) +
ggtitle('16S: Shared incorrect species') +
ylab('Species')
c
a + b + c & ylim(c(0, 20)) &
theme(
# Hide panel borders and remove grid lines
panel.border = element_blank(),
panel.grid.major = element_blank(),
panel.background = element_blank(),
panel.grid.minor = element_blank(),
#panel.grid.major.y = element_line(),
# Change axis line
axis.line = element_line(colour = "black")
)
16S/12S relative plots
sixteenS_relative_plot / twelve_s_relative_plot
Let’s make without titles, but with a/b
(sixteenS_relative_plot + ggtitle('') + ylab(''))/ (twelve_s_relative_plot + ggtitle('')) +
plot_annotation(tag_levels = c('A','B')) +
plot_layout(guides = 'collect')
(sixteenS_score_plot +geom_point() + theme(axis.title.x = element_blank()))/ (twelveS_scoreS_plot + geom_point())
12S exclusion databases
twelve_exclusions <- data |> filter(str_starts(Subject, '12s_v010_final.fasta_Taxonomies.CountedFams.txt_')) |>
filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa')
table(twelve_exclusions$Subject)##
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 1242
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 1248
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 1259
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 1234
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 1213
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 1244
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 1232
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 1230
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 1267
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 1246
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 1274
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 1265
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 1196
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 1213
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 1276
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 1296
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 1250
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 1299
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 1257
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 1266
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 1190
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 1179
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 1170
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 1166
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 1146
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 1193
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 1159
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 1157
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 1183
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 1216twelve_exclusions_split <- twelve_exclusions |> separate(Subject, into = c('before', 'hit'), sep='.txt_') |>
# get rid of leftover non-subsetted databases
filter(!is.na(hit)) |>
separate(hit, into=c('Database', 'after'), sep='_subset')twelve_exclusions_split_averaged <- twelve_exclusions_split |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type, Database, after) |>
summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
FN = sum(is.na(species) & !is.na(True_species))) |>
mutate(sums = TP + FP + TN + FN) |>
mutate(missing = 102 - sums) |>
mutate(FN = FN + missing) |>
mutate(sums = TP + FP + TN + FN) |>
select(-c(missing, sums)) |>
group_by(Type, Database) |>
summarise(mean_TP = mean(TP),
mean_FP = mean(FP),
mean_TN = mean(TN),
mean_FN = mean(FN)) |>
rowwise() |>
mutate(recall = recall(mean_TP, mean_FN),
precision = precision(mean_TP, mean_FP),
f1 = f1(precision, recall),
f0.5 = f0.5(precision, recall),
accuracy = accuracy(mean_TP, mean_FP, mean_FN, mean_TN)) ## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
## `summarise()` has grouped output by 'Type'. You can override using the
## `.groups` argument.twelve_exclusions_split_averaged <- twelve_exclusions_split_averaged |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database))f1_pl <- twelve_exclusions_split_averaged |>
ggplot(aes(x = Type, y = f1, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()f0.5_pl <- twelve_exclusions_split_averaged |>
ggplot(aes(x = Type, y = f0.5, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()precision_pl <- twelve_exclusions_split_averaged |>
ggplot(aes(x = Type, y = precision, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()recall_pl <- twelve_exclusions_split_averaged |>
ggplot(aes(x = Type, y = recall, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()(f1_pl / f0.5_pl / precision_pl / recall_pl) + plot_layout(guides = 'collect')
Lets zero in on the precision and make boxplots with jitter dots
un_summarised_twelve <- twelve_exclusions_split |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type, Database, after) |>
summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
FN = sum(is.na(species) & !is.na(True_species))) |>
mutate(sums = TP + FP + TN + FN) |>
mutate(missing = 102 - sums) |>
mutate(FN = FN + missing) |>
mutate(sums = TP + FP + TN + FN) |>
select(-c(missing, sums)) |>
rowwise() |>
mutate(recall = recall(TP, FN),
precision = precision(TP, FP),
f1 = f1(precision, recall),
f0.5 = f0.5(precision, recall),
accuracy = accuracy(TP, FP, FN, TN)) |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database))## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.un_summarised_twelve |> group_by(Type, Database) |> mutate(best = max(mean(precision))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = precision)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('Precision') +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_twelve |> group_by(Type, Database) |> mutate(best = max(mean(f0.5))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = f0.5)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('f0.5') +
ylim(c(0, 1)) +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_twelve |> group_by(Type, Database) |> mutate(best = max(mean(recall))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = recall)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('f0.5') +
ylim(c(0, 1)) +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_twelve |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Qiime2', 'TNT')) |>
ggplot(aes(x=Database, y = precision, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot() +
labs(fill='Type') +
ylab('Precision') +
theme_minimal()
false_positives <- un_summarised_twelve |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |>
ggplot(aes(x=Database, y = FP/102*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot(outlier.shape=NA) +
labs(fill='Type') +
ylab('False positives (%)') +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE)+
theme_minimal()
false_positives
true_positives <- un_summarised_twelve |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |>
ggplot(aes(x=Database, y = TP/102*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot(outlier.shape=NA) +
labs(fill='Type') +
ylab('True positives (%)') +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE)+
theme_minimal()
true_positives
false_positives/ true_positives + plot_layout(guides = 'collect') & coord_flip()
### Phylogenetic diversity
We can also easily calculate alpha diversity across these tools, as alpha diversity is the number of species. We treat classifiers/Types as sites.
spec_summarised <- twelve_exclusions_split |>
group_by(Type, Query, Database, after) |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database)) |>
filter(!is.na(species)) |>
summarise(`Alpha diversity index` = length(unique(species)))## `summarise()` has grouped output by 'Type', 'Query', 'Database'. You can
## override using the `.groups` argument.spec_summarised |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot() +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) + coord_flip() + theme_minimal()
Let’s also do not all of the classifiers
spec_summarised |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot() +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) + coord_flip() + theme_minimal()
a <- spec_summarised |>
filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0','Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
group_by(Database) |>
arrange(Database) |>
group_map(~aov(`Alpha diversity index` ~ Type, data=.))
names(a) <- spec_summarised |> arrange(Database) |> pull(Database) |> unique() # I don't like this :(
a## $`30%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 4149.933 1070.800
## Deg. of Freedom 5 54
##
## Residual standard error: 4.453047
## Estimated effects may be unbalanced
##
## $`50%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 3674.483 653.700
## Deg. of Freedom 5 54
##
## Residual standard error: 3.479304
## Estimated effects may be unbalanced
##
## $`70%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 2567.749 805.975
## Deg. of Freedom 5 52
##
## Residual standard error: 3.936943
## Estimated effects may be unbalancedlibrary(agricolae)
groupslist <- list()
for(key in names(a)) {
print(key)
groupslist[[key]] <- HSD.test(a[[key]], 'Type')$groups|>
as_tibble(rownames = 'Type') |>
select(-`Alpha diversity index`)
}## [1] "30%"
## [1] "50%"
## [1] "70%"groups_df <- bind_rows(groupslist, .id='Database')spec_summarised |>
filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
left_join(groups_df, by = c('Database', 'Type')) |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot(outlier.shape=NA) +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) +
geom_text(aes(x = Type, y = max(`Alpha diversity index`) + 2, label = groups),
#col = 'black',
size = 4) +
#coord_flip() +
theme_minimal() +
theme(axis.text.x = element_text( angle = 90, hjust = 1)) +
guides(fill="none")
16S exclusion databases
sixteen_exclusions <- data |> filter(str_starts(Subject, '16S_v04_final.fasta_Taxonomies.')) |>
filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa')
table(sixteen_exclusions$Subject)##
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta
## 1269
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta
## 1313
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta
## 1394
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta
## 1290
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta
## 1320
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta
## 1339
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta
## 1348
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta
## 1321
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta
## 1263
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta
## 1287
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta
## 1411
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta
## 1347
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta
## 1401
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta
## 1334
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta
## 1314
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta
## 1442
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta
## 1380
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta
## 1395
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta
## 1422
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta
## 1423
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta
## 1263
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta
## 1288
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta
## 1283
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta
## 1267
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta
## 1334
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta
## 1274
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta
## 1234
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta
## 1319
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta
## 1262
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta
## 1274sixteen_exclusions_split <- sixteen_exclusions |> separate(Subject, into = c('before', 'hit'), sep='.txt_') |>
# get rid of leftover non-subsetted databases
filter(!is.na(hit)) |>
separate(hit, into=c('Database', 'after'), sep='_subset')sixteen_exclusions_split_averaged <- sixteen_exclusions_split |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type, Database, after) |>
summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
FN = sum(is.na(species) & !is.na(True_species))) |>
mutate(sums = TP + FP + TN + FN) |>
mutate(missing = 102 - sums) |>
mutate(FN = FN + missing) |>
mutate(sums = TP + FP + TN + FN) |>
select(-c(missing, sums)) |>
group_by(Type, Database) |>
summarise(mean_TP = mean(TP),
mean_FP = mean(FP),
mean_TN = mean(TN),
mean_FN = mean(FN)) |>
rowwise() |>
mutate(recall = recall(mean_TP, mean_FN),
precision = precision(mean_TP, mean_FP),
f1 = f1(precision, recall),
f0.5 = f0.5(precision, recall),
accuracy = accuracy(mean_TP, mean_FP, mean_FN, mean_TN)) ## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
## `summarise()` has grouped output by 'Type'. You can override using the
## `.groups` argument.sixteen_exclusions_split_averaged <- sixteen_exclusions_split_averaged |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database))f1_pl <- sixteen_exclusions_split_averaged |>
ggplot(aes(x = Type, y = f1, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()f0.5_pl <- sixteen_exclusions_split_averaged |>
ggplot(aes(x = Type, y = f0.5, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()precision_pl <- sixteen_exclusions_split_averaged |>
ggplot(aes(x = Type, y = precision, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()recall_pl <- sixteen_exclusions_split_averaged |>
ggplot(aes(x = Type, y = recall, group = Database, color = Database, fill = Database)) +
geom_point() +
geom_line() +
theme_minimal()(f1_pl / f0.5_pl / precision_pl / recall_pl) + plot_layout(guides = 'collect')
Lets zero in on the precision and make boxplots with jitter dots
un_summarised_sixteen <- sixteen_exclusions_split |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|>
mutate(species = na_if(species, 'dropped')) |>
mutate(genus = na_if(genus, 'dropped')) |>
mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
!is.na(species) & True_species != species ~ 'Incorrect species',
!is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
!is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
!is.na(family) & True_family == family ~ 'Correct family',
!is.na(family) & True_family != family ~ 'Incorrect family',
TRUE ~ NA)) |>
group_by(Type, Database, after) |>
summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
FN = sum(is.na(species) & !is.na(True_species))) |>
mutate(sums = TP + FP + TN + FN) |>
mutate(missing = 112 - sums) |>
mutate(FN = FN + missing) |>
mutate(sums = TP + FP + TN + FN) |>
select(-c(missing, sums)) |>
rowwise() |>
mutate(recall = recall(TP, FN),
precision = precision(TP, FP),
f1 = f1(precision, recall),
f0.5 = f0.5(precision, recall),
accuracy = accuracy(TP, FP, FN, TN)) |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database))## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.un_summarised_sixteen |> group_by(Type, Database) |> mutate(best = max(mean(precision, na.rm = TRUE))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = precision)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('Precision') +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_sixteen |> group_by(Type, Database) |> mutate(best = max(mean(f0.5, na.rm=TRUE))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = f0.5)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('f0.5') +
ylim(c(0, 1)) +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_sixteen |> group_by(Type, Database) |> mutate(best = max(mean(recall))) |>
ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = recall)) +
geom_boxplot(outlier.shape = NA) +
coord_flip() +
theme_minimal() +
xlab('Type') +
ylab('f0.5') +
ylim(c(0, 1)) +
geom_point(position = position_jitterdodge(), alpha=0.5)
un_summarised_sixteen |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Qiime2', 'TNT')) |>
ggplot(aes(x=Database, y = precision, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot() +
labs(fill='Type') +
ylab('Precision') +
theme_minimal()
false_positives <- un_summarised_sixteen |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |>
ggplot(aes(x=Database, y = FP/112*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot(outlier.shape=NA) +
labs(fill='Type') +
ylab('False positives (%)') +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE)+
theme_minimal()
false_positives
true_positives <- un_summarised_sixteen |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |>
ggplot(aes(x=Database, y = TP/112*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) +
geom_boxplot(outlier.shape=NA) +
labs(fill='Type') +
ylab('True positives (%)') +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE)+
theme_minimal()
true_positives
false_positives/ true_positives + plot_layout(guides = 'collect') & coord_flip()
### Phylogenetic diversity
We can also easily calculate alpha diversity across these tools, as alpha diversity is the number of species. We treat classifiers/Types as sites.
spec_summarised <- sixteen_exclusions_split |>
group_by(Type, Query, Database, after) |>
mutate(Database = case_when ( Database == 'fifty' ~ '50%',
Database == 'seventy' ~ '70%',
Database == 'thirty' ~ '30%',
TRUE ~ Database)) |>
filter(!is.na(species)) |>
summarise(`Alpha diversity index` = length(unique(species)))## `summarise()` has grouped output by 'Type', 'Query', 'Database'. You can
## override using the `.groups` argument.spec_summarised |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot() +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) + coord_flip() + theme_minimal()
Let’s also do not all of the classifiers
spec_summarised |>
filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot() +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) + coord_flip() + theme_minimal()
a <- spec_summarised |>
filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
group_by(Database) |>
arrange(Database) |>
group_map(~aov(`Alpha diversity index` ~ Type, data=.))
names(a) <- spec_summarised |> arrange(Database) |> pull(Database) |> unique() # I don't like this :(
a## $`30%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 8446.646 1729.975
## Deg. of Freedom 5 52
##
## Residual standard error: 5.767907
## Estimated effects may be unbalanced
##
## $`50%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 6966.745 3111.000
## Deg. of Freedom 5 49
##
## Residual standard error: 7.968048
## Estimated effects may be unbalanced
##
## $`70%`
## Call:
## aov(formula = `Alpha diversity index` ~ Type, data = .)
##
## Terms:
## Type Residuals
## Sum of Squares 4175.647 788.914
## Deg. of Freedom 5 51
##
## Residual standard error: 3.933053
## Estimated effects may be unbalancedlibrary(agricolae)
groupslist <- list()
for(key in names(a)) {
print(key)
groupslist[[key]] <- HSD.test(a[[key]], 'Type')$groups|>
as_tibble(rownames = 'Type') |>
select(-`Alpha diversity index`)
}## [1] "30%"
## [1] "50%"
## [1] "70%"groups_df <- bind_rows(groupslist, .id='Database')spec_summarised |>
filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |>
left_join(groups_df, by = c('Database', 'Type')) |>
ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) +
geom_boxplot(outlier.shape=NA) +
geom_point(aes(color=Type),
position = position_jitterdodge(jitter.width = 0.2),
alpha=0.5,
show.legend = FALSE) +
facet_wrap(~Database) +
geom_text(aes(x = Type, y = max(`Alpha diversity index`) + 2, label = groups),
#col = 'black',
size = 4) +
#coord_flip() +
theme_minimal() +
theme(axis.text.x = element_text( angle = 90, hjust = 1)) +
guides(fill="none")